抗氧化剂吡咯烷二硫代氨基甲酸盐对宫颈癌SiHa细胞增殖和凋亡的影响

Effect of antioxidant PDTC on the proliferation and apoptosis of human cervical carcinoma SiHa cell lines in vitro

目的:探讨抗氧化剂吡咯烷二硫代氨基甲酸盐(PDTC)的体外抗宫颈癌作用机制。方法:体外培养SiHa细胞,PDTC处理后采用WST-8方法检测SiHa细胞的增殖活性;Hoechst 33258染色、Annexin Ⅴ-FITC/PI双标法细胞凋亡试剂盒检测细胞凋亡;凝胶迁移变动分析(EMSA)检测SiHa细胞核内核转录因子-κB(NF-κB)活性。结果:PDTC能有效抑制SiHa细胞增殖和诱导SiHa细胞凋亡;染色可见典型的凋亡细胞形态;各组间早期凋亡率比较差异有显著性(P<0.01),100μmol/LPDTC晚期凋亡率较其他组明显升高(P<0.01);PDTC可抑制SiHa细胞核内NF-κB活性,其抑制强度与作用24h细胞生长抑制率呈直线正相关(r=0.931,P=0.035),与作用48h早期凋亡率呈曲线正相关(r=1.000,P=0.011)。结论:PDTC抑制SiHa细胞增殖与其诱导细胞凋亡作用有关,而致凋亡作用可能与其抑制NF-κB信号转导通路,启动凋亡程序有关。

Objective To investigate the effect of antioxidant pyrrolidinedithiocarbamate（PDTC） on human cervical carcinoma SiHa cell lines.Methods Human cervical carcinoma SiHa cell lines which were cultured in vitro were treated with different concentrations of PDTC for different durations.Then the proliferation of SiHa cells were detected by WST-8 assay,the apoptosis was assessed by Hoechst 33258 staining and Annexin Ⅴ-FITC /PI double-labeled apoptosis detection kit,and the activity of nuclear factor-κB was determined by electrophoretic mobility shift assay（EMSA）.Results PDTC could significantly inhibit the proliferation and induce the apoptosis of SiHa cells in a concentration-dependent manner and in partly time-dependent manner.Apoptosis could be observed by Hoechst 33258 staining.After being treated with 20,50,and 100 μmol /L PDTC for 48 hours,the early apoptosis rates of SiHa cells were（7.00 ± 1.50）%,（14.22 ± 4.02）%,and（34.39 ± 4.34）%,respectively（P〈 0.01）.The late apoptosis rate of SiHa cells treated with 100 μmol /L PDTC was（12.16 ± 1.54）%,which was significantly higher than those in other groups（P〈0.01）.Furthermore,EMSA revealed that PDTC could significantly and effectively decrease constitutive NF-κB DNA binding activity at max for 4 h and in a dose-dependent manner.The inhibitory effects showed a linear positive correlation to the proliferation inhibitory rate at 24 h（r = 0.931,P = 0.035） and to a guadratic positive corelation the early apoptosis rate at 48 h（r = 1.000,P = 0.011）.Conclusion PDTC can inhibit NF-κB signaling pathway,so as to induce the apoptosis of SiHa cells and finally inhibit the proliferation of them.